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US20140194513A1 - Fermented vegetable oil and composition including same - Google Patents

Fermented vegetable oil and composition including same Download PDF

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Publication number
US20140194513A1
US20140194513A1 US14/241,137 US201214241137A US2014194513A1 US 20140194513 A1 US20140194513 A1 US 20140194513A1 US 201214241137 A US201214241137 A US 201214241137A US 2014194513 A1 US2014194513 A1 US 2014194513A1
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Prior art keywords
fermented
oil
vegetable oil
bacteria
fatty acid
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US14/241,137
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English (en)
Inventor
Kwang-Nyeon Kim
Ju-Hyun Son
Hee-Sik Kim
Jong-Seok Yun
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Korea Research Institute of Bioscience and Biotechnology KRIBB
Damy Chemical Co Ltd
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Damy Chemical Co Ltd
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Assigned to DAMY CHEMICAL CO., LTD. reassignment DAMY CHEMICAL CO., LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: KIM, KWANG-NYEON, SON, JU-HYUN, YUN, JONG SEOK
Assigned to KOREA RESEARCH INSTITUTE OF BIOSCIENCE AND BIOTECHNOLOGY reassignment KOREA RESEARCH INSTITUTE OF BIOSCIENCE AND BIOTECHNOLOGY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: KIM, HEE-SIK
Publication of US20140194513A1 publication Critical patent/US20140194513A1/en
Abandoned legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
    • A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/361—Carboxylic acids having more than seven carbon atoms in an unbroken chain; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings or cooking oils characterised by the production or working-up
    • A23D9/04—Working-up
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • C12P7/6431—Linoleic acids [18:2[n-6]]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine

Definitions

  • the present invention relates to a fermented vegetable oil and a composition including same, and more particularly, and provides a fermented vegetable oil, which has the effects of enhancing emulsion stability due to water retention ability, improving texture and flavor and enhancing moisturization, according to being fermented by bacteria.
  • a fermented oil refers to an oil, which is prepared by fermenting oriental medicine such as ginseng, extracting fat-soluble ingredients therefrom and adding thereof to an oil, or which is prepared by adding oriental medicine such as ginseng to an oil followed by maturing thereof.
  • an oil which is prepared by fermenting fishes or coconut and extracting fat-soluble ingredients therefrom, or an oil, which is prepared by soaking germinating or steaming oil seeds (soy bean, sun flower seed, grape seed, sesame seed and the like) and then extracting an oil therefrom, is also called a fermented oil.
  • the present invention provides a fermented vegetable oil, which has the effects of enhancing emulsion stability due to water retention ability, improving texture and flavor, and enhancing moisturizing capacity, according to being fermented by bacteria.
  • the present invention provides a method for preparing a fermented vegetable oil, which is fermented by bacteria by a simple method.
  • the present invention provides a composition for skin external preparations or cosmetics, which has the effect of enhancing emulsion stability, improving texture and flavor, and enhancing moisturizing capacity, by containing a fermented vegetable oil fermented by bacteria as an active ingredient.
  • the fermented vegetable oil of the present invention is characterized by being fermented by bacteria.
  • the method for preparing the fermented vegetable oil of the present invention is characterized by comprising: (a) a step of firstly culturing bacteria in a culture solution at an aerobic condition; (b) a step of secondarily culturing after adding a vegetable oil to the culture solution; and (c) a step of collecting the secondarily cultured vegetable oil.
  • the composition for skin external preparations or cosmetics of the present invention is characterized by comprising the fermented vegetable oil, which is fermented by bacteria, as an active ingredient.
  • the fermented vegetable oil according to the present invention is excellent on emulsion stability due to water retention ability. Accordingly, it is easy to manufacture a toner or essence-type products by applying the oil thereto.
  • the fermented vegetable oil according to the present invention has lighter texture and more excellent moisture feeling than before fermentation.
  • the fermented vegetable oil according to the present invention has higher stability and usability than a natural oil and a matured oil having similar effects.
  • FIG. 1 the analysis result of the emulsion stability, wherein FIG. 1 a is the result of measuring the emulsion activity of an olive oil, FIG. 1 b is an image of estimating the emulsion stability of an olive oil depending on time, FIG. 1 c is the result of measuring the emulsion activity of a fermented olive oil, FIG. 1 d is an image of estimating the emulsion stability of a fermented olive oil depending on time, FIG. 1 e is the result of comparing the emulsion activities of fermented oils depending on type, and FIG. 1 f to FIG. 1 m are images of estimating the emulsion activities of fermented oils depending on type.
  • FIG. 2 the result of analyzing fatty acid by gas chromatography, wherein FIG. 2 a is a fatty acid distribution chart, FIG. 2 b is the result of analyzing fatty acid of an olive oil, and FIG. 2 c is the result of analyzing fatty acid of a fermented olive oil.
  • FIG. 3 the result of measuring the acid value, wherein FIG. 3 a is the result of measuring the acid value of an olive oil, and FIG. 3 b is the result of measuring the acid value of a fermented olive oil.
  • FIG. 4 the result of measuring the influence on the cell growth and toxicity in human fibroblasts, wherein FIG. 4 a is the result of the cell viability of a soy bean oil and a fermented soy bean oil, FIG. 4 b is the result of the cell viability of an olive oil and a fermented olive oil, FIG. 4 c is the result of the cell viability of a green tea oil and a fermented green tea oil, and FIG. 4 d is the result of the cell viability of an argan oil and a fermented argan oil.
  • FIG. 5 the result of analyzing ingredients by using TLC (Thin layer chromatography), wherein FIG. 5 a is the result of 2D-TLC of an olive oil, a fermented olive oil, linoleic acid and linolenic acid, and FIG. 5 b is the result of 2D-TLC of a soy bean oil, a fermented soy bean oil, a green tea oil, a fermented green tea oil, an argan oil and a fermented argan oil.
  • TLC Thin layer chromatography
  • the fermented vegetable oil of the present invention is characterized by being fermented by bacteria.
  • the bacteria may be SY16 (KCTC 8950P) of Pseudozima sp., preferably, and the SY16 (KCTC 8950P) of Pseudozima sp. is in the shape of cylinder and grows in polar germination mode, and forms endospores, which differ from hyphae and ascospores.
  • the fermented vegetable oil of the present invention is characterized that it is fermented by the above bacteria in a medium containing a vegetable oil having the acid value of 0.100 ⁇ 2.000.
  • the vegetable oil fermented in the medium may be any general vegetable oil such as olive oil, soy bean oil, green tea oil and argan oil, and it may be an oil having the acid value of 0.100 ⁇ 2.000, preferably.
  • the acid value is preferred to be 0.1 ⁇ 0.5.
  • the fermented vegetable oil of the present invention is characterized by having higher essential fatty acid content than before fermentation, by being fermented by the bacteria. At this time, the essential fatty acid content after fermentation is 5.0 ⁇ 140.0 times higher than before fermentation.
  • the essential fatty acid is preferred to be linoleic acid.
  • the linoleic acid is a key component of the epidermal lipid bilayer, and is contained in ceramides of the epidermal keratin layer. This linoleic acid is effective in skin moisturizing, skin homeostasis maintenance, formation and protection of the skin permeation layer.
  • the essential fatty acid contained in the fermented vegetable oil of the present invention may be linoleic acid, preferably.
  • the fermented vegetable oil of the present invention is characterized by having higher free fatty acid content than before fermentation, by being fermented by the bacteria. At this time, the free fatty acid content after fermentation is 5.0 ⁇ 140.0 times higher than before fermentation.
  • the free fatty acid refers to fatty acid that is in unbound glyceride form.
  • texture becomes improved because stickiness is reduced, and it is effective in enhancing oiliness after use. Accordingly, it is preferred that the fermented vegetable oil of the present invention has higher free fatty acid content.
  • the method for preparing the fermented vegetable oil of the present invention includes (a) a step of firstly culturing bacteria in a culture solution at an aerobic condition; (b) a step of secondarily culturing after adding a vegetable oil to the culture solution; and (c) a step of collecting the secondarily cultured vegetable oil.
  • bacteria are firstly cultured in a culture solution at an aerobic condition.
  • the bacteria may be SY16 (KCTC 8950P) of Pseudozima sp., preferably.
  • the SY16 (KCTC 8950P) of Pseudozima sp. is in the shape of cylinder and grows in polar germination mode, and forms endospores, which differ from hyphae and ascospores.
  • the culture solution does not contain a vegetable oil yet, and it is very important to mix various kinds of culture materials to make the vegetable oil be fermented well.
  • the first culture time may be 24 ⁇ 72 hours, and 30 ⁇ 60 hours, preferably because the lipase activity is the highest within the said range.
  • the optimized culture condition is determined by the acid value of a vegetable oil.
  • the acid value of a vegetable oil may be in the range of 0.100 ⁇ 2.000, preferably, and it may be in the range of 0.1 ⁇ 0.5, more preferably, because the lower acid value makes the higher fermentation efficiency.
  • the vegetable oil is added in an amount of 100 parts by weight or less, based on 100 parts by weight of the above culture solution, in which the vegetable oil is not added yet.
  • the amount of the vegetable oil added may be properly adjusted according to the desired amount of a fermented vegetable oil, but when the vegetable oil is added in an amount of over 100 parts by weight, based on the 100 parts by weight of the culture solution containing the vegetable oil, it is concerned that the oil may not be fermented enough.
  • the second culture time may be 72 ⁇ 120 hours, preferably because the time enough to produce the maximum amount of free fatty acid should be secured as the fermentation, resulting from the secondary culture, goes on. Further, the time may vary depending on the kinds of vegetable oils. Accordingly, the secondary culture may be finished when the amount of the free fatty acid is not increased any more after measuring the amount of the free fatty acid increased with time.
  • the collected vegetable oil is a fermented oil, and it can be used as an active ingredient of a composition for skin external preparations or cosmetics.
  • the present invention includes a composition, which contains the fermented vegetable oil fermented by bacteria, as an active ingredient, and is used for skin external preparations or cosmetics.
  • the inventive fermented vegetable oil which is fermented by bacteria, is suitable for a composition for skin external preparations or cosmetics because it has excellent texture and moisturizing capacity due to its high emulsion activity and emulsion stability, and much higher contents of essential fatty acid and free fatty acid than general oils.
  • the amount of the fermented vegetable oil contained in the composition for skin external preparations and cosmetics may be 2 ⁇ 40 wt %, preferably.
  • the amount of the fermented vegetable oil is less than 2 wt %, the effect of the product may be meager because the fermented vegetable oil is not contained enough, and if the amount is over 40 wt %, it may be difficult to be formulated.
  • compositions for skin external preparations and cosmetics of the present invention may contain general ingredients used for compositions for skin external preparations and cosmetics in addition to the said fermented vegetable oil, for example, general additives such as antioxidant, stabilizer, solubilizer, vitamin, dye and fragrance, and carriers.
  • general additives such as antioxidant, stabilizer, solubilizer, vitamin, dye and fragrance, and carriers.
  • composition for skin external preparations and cosmetics of the present invention may be formulated in a wide variety of forms commonly used in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleanser, oil, powder foundation, emulsion foundation, wax foundation and spray, but not limited thereto.
  • the carrier contained in the present composition for skin external preparations and cosmetics may be any carrier commonly used in the art depending on the type of the formulation.
  • the carrier ingredient may be animal and vegetable fats, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and the like.
  • the carrier ingredient may be lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder.
  • the spray may further comprise propellants such as chlorofluorohydrocarbon, propane/butane or dimethyl ether.
  • the carrier ingredient may be solvent, solubilizer and emulsifier.
  • it may be water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, in particular, cottonseed oil, peanut oil, maize germ oil, olive oil, castor oil and sesame seed oil, glycerol fatty esters, polyethylene glycol and fatty acid esters of sorbitan.
  • the carrier ingredient may be liquid diluents, for example, water, ethanol or propylene glycol, suspending agents, for example, ethoxylated isosteary alcohol, polyoxyethylene sorbitol ester and poly oxyethylene sorbitan ester, micocrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth.
  • suspending agents for example, ethoxylated isosteary alcohol, polyoxyethylene sorbitol ester and poly oxyethylene sorbitan ester, micocrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth.
  • the carrier ingredient may be aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glycerides, fatty acid diethanolamide, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid ester.
  • the carrier ingredient may be alkali metal salts of fatty acids, salts of fatty acid hemiesters, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohol, vegetable oil, glycerol, sugars and the like.
  • the cosmetic composition of the present invention may include other additives in addition to the carrier, for example, preservative, antioxidant, stabilizer, solubilizer, vitamin, dye and fragrance.
  • SY16 [KCTC 8950P] of Pseudozima sp. was inoculated to each 1 L of the following 8 medium compositions, and cultured at 25° C., 300 rpm, under an aerobic condition.
  • composition 1 Composition 2
  • Composition 3 Composition 4 Glucose 15 g/L Sucrose 10 g/L Glucose 10 g/L Glucose 10 g/L Olive oil 100 g/L Olive oil 100 g/L Olive oil 100 g/L (NH 4 ) 2 SO 4 1 g/L Yeast extract 3 g/L Yeast extract 3 g/L Yeast extract 3 g/L K 2 HPO 4 2.5 g/L Skim milk 3 g/L Malt extract 3 g/L Malt extract 3 g/L Malt extract 3 g/L MgSO 4 0.5 g/L Casein 3 g/L Peptone 5 g/L Peptone 3 g/L CaCl 2 0.1 g/L Soya bean meal Distilled water Soya bean meal MnSO 4 0.02 g/L 3 g/L 879 g/L 3 g/L NaH 2 PO 4
  • the bacterial growth was measured by culturing the bacteria at each condition, centrifuging thereof to remove an oil layer, strongly mixing the resulting solution for 20 sec, diluting the mixed solution 10 times with distilled water, and then measuring absorbance (660 nm) of each sample.
  • the culture solution was centrifuged at 12,000 rpm for 3 min, the supernatant was used as a crude enzyme solution, and then the lipase activity was measured by microplate analyzing method measuring the amount of p-nitrophenol (pNP) bound to a substrate, which is released by the said enzyme, as the change of absorbance (405 nm).
  • the composition 4 showed the highest growth and lipase activity. Accordingly, the composition was used for preparing the next fermented oil.
  • the amount that the crude enzyme 100 ⁇ L produces tyrosine 1 ⁇ g for 1 min was used as 1 unit.
  • the composition 4 showed the highest growth and lipase activity of the bacteria. Accordingly, the composition 4 was determined as the optimum medium composition for the future fermented oil production by bacteria.
  • SY16 [KCTC 8950P] of Pseudozima sp. was inoculated to a culture solution (glucose 10 g/L, olive oil 100 g/L, yeast extract 3 g/L, malt extract 3 g/L, peptone 3 g/L, soya bean meal 3 g/L, (NH 4 ) 2 SO 4 2 g/L, KH 2 PO 4 1 g/L, MgSO 4 0.5 g/L, CaCl 2 0.1 g/L, NaCl 0.1 g/L: the medium of the composition 4) 1 L, and cultured at 25° C., 300 rpm under an aerobic condition.
  • a culture solution glucose 10 g/L, olive oil 100 g/L, yeast extract 3 g/L, malt extract 3 g/L, peptone 3 g/L, soya bean meal 3 g/L, (NH 4 ) 2 SO 4 2 g/L, KH 2 PO 4 1
  • the culture solution 1 ml was collected at each time point (3, 7, 12, 24, 48, 72 and 96 hr), and then the growth and lipase activity of the bacteria were measured at 660 nm by using a spectrophotometer. As the result of measuring the lipase activity, it was found that the first culture time showing the highest lipase activity was 48 hr.
  • SY16 [KCTC 8950P] of Pseudozima sp. was inoculated to the culture solution as described above 100 L, and cultured for 48 hours for conducting the first culture. Then, olive oil 100 L was added to the culture solution, and secondarily cultured at 25° C., 500 rpm under an aerobic condition. After the second culture, the culture solution 0.3 ⁇ 0.5 L was collected at each time point (3, 7, 12, 24, 48, 72, 96 and 120 hr), and then only oil part was collected. The degree of oil fermentation was checked by measuring the amount of free fatty acid in the collected oil. As the result of measuring the increased amount of the free fatty acid, the amount of the free fatty acid was the highest at the time of 96 hr after adding the oil, but the amount was not increased after that time.
  • Emulsion stability means the ability of an emulsifier, which stabilizes emulsion after forming the emulsion or at the condition of mixing, high temperature, centrifugation and the like.
  • each 1 ml of an olive oil and a fermented olive oil was dissolved in each 10 ml of buffer solutions depending on pH, and emulsified by completely stirring thereof for 2 min. Then, the resulting solution was kept for 10 min, and then the emulsion activity was measured by measuring absorbance at 620 nm.
  • the emulsion stability was obtained as the percentage of the absorbance after 24 hrs to the initial absorbance. At pH 9.0, the absorbance was beyond the maximum absorbance value of the fermented oil, but the value set by a prediction program of the device was recorded.
  • the emulsion activities of the olive oil and the fermented olive oil depending on pH were listed in the following Table 2.
  • the emulsion activity of the fermented olive oil was rapidly increased from pH 5.0.
  • the emulsion activity at pH 9.0 was 50% or higher than at pH 5.0.
  • the emulsion activity of the olive oil was increased at pH 9.0, but the emulsion activity at pH 9.0 was below the emulsion activity of the fermented olive oil at pH 5.0.
  • the emulsion activity of the fermented olive oil was showed from pH 2.0, and it was confirmed that the emulsion activity of the fermented olive oil at pH 9.0 was 2.5 times or higher than that of the olive oil.
  • FIG. 1 a is the result of measuring the emulsion activity of the olive oil
  • FIG. 1 c is the result of measuring the emulsion activity of the fermented olive oil.
  • FIG. 1 b is the image about the olive oil
  • FIG. 1 d is the image about the fermented olive oil.
  • the olive oil After 24 hrs, the olive oil showed very little emulsion activity, but the fermented olive oil showed excellent emulsion activity as much as 4 times or more than the olive oil.
  • FIG. 1 e is the result of measuring the emulsion activities of the fermented oils
  • FIG. 1 f to FIG. 1 m are images about the emulsion activities.
  • the fermented oils have improved emulsifying capacity because they showed at least 2 times higher emulsion activities than before fermentation.
  • the fermented olive oil had better moisture feeling, lower stickiness and not oiliness, compared with the olive oil. Accordingly, it could be found that the fermented oil of the present invention has excellent skin texture such as moisture feeling, stickiness, oiliness and the like.
  • Fatty acid methyl ester was deacylated, the fatty acid, which can be dissolved in hexane, was prepared as boron trifluoride methanol mixture, and then, analyzed by GC analyzer (JMS-SX 102A, JEOL, Tokyo, Japan).
  • FIG. 2 a is a distribution chart of fatty acid
  • FIG. 2 b is the result of analyzing fatty acid in the olive oil
  • FIG. 2 c is the result of analyzing fatty acid in the fermented olive oil
  • FIG. 2 c is the result of analyzing fatty acid in the green tea oil
  • FIG. 2 d is the result about the fermented green tea oil.
  • the fatty acid composition of the olive oil and the fermented olive oil As the result of comparing the fatty acid composition of the olive oil and the fermented olive oil, it was confirmed that the content of essential fatty acid such as linoleic acid was more increased in the fermented olive oil than in the olive oil. Further, as the result of comparing the fatty acid composition of the green tea oil and the fermented green tea oil, it was confirmed that the essential fatty acid content such as linoleic acid was more increased in the fermented green tea oil than in the green tea oil.
  • the fermented oil improved water retention ability and absorbing ability of the skin as the essential fatty acid content was increased. Accordingly, it was confirmed that the fermented oil is effective for improving moisture feeling.
  • the acid value is the amount of free fatty acid that is in unbound glyceride form.
  • the acid value was measured by using a metrohm titrator.
  • a sample was added in an amount to make the estimated consumption of a titrant (KOH 0.05 mol/L) is between 1 ml and 10 ml.
  • MET mode of the device was selected, U (voltage) was selected among measuring values, MET U mode was loaded, an electrode and a burette tip were set in a beaker, and then the test was started.
  • FIG. 3 a is the acid value of the olive oil
  • FIG. 3 b is the acid value of the fermented olive oil.
  • the acid value of the olive oil was 0.1294 mgKOH/g and that of the fermented olive oil was 8.1022 mgKOH/g. Accordingly, it was confirmed that the amount of the free fatty acid of the fermented olive oil was increased 50 times or higher than that of the olive oil.
  • the olive oil and the fermented olive oil were treated to the cultured human fibroblast at various concentrations, and then cell viabilities were compared for verifying influence of the fermentation process on the cell growth and toxicity.
  • MTT is absorbed into cells, and forms formazan in mitochondria by succinic acid dehydrogenase. Intracellular accumulation of the formazan means the activity of mitochondria, broadly, the cell activity. Accordingly, the cell growth and toxicity can be tested.
  • Olive oil and fermented olive oil were treated to cells cultured in a 96 well plate at the concentration of 0.5% ⁇ 0.0019%, respectively, and then cultured for 48 hours in a 5% CO 2 , 37° C. incubator. The cells were reacted in MTT solution for 3-4 hours, and then O.D value was measured at 540 nm by ELISA analyzer.
  • the cell viability may be calculated by the following formula.
  • DMSO-treated group was used as a control group, and the results of tests conducted by adding soy bean oil, green tea oil and argan oil before fermentation, and soy bean oil, green tea oil and argan oil after fermentation were shown in FIG. 4 .
  • An olive oil and a fermented olive oil as a sample were diluted to a certain concentration (1/10, 1/5). 2-3 drops of each sample were dropped on Silica TLC, and analyzed at solvent condition having good resolution to find the optimum solvent condition.
  • the TLC with the sample was developed with a solvent having good resolution as a developing solvent in a chamber. The developed TLC was picked out and dried. The TLC was dipped in 10% sulfuric acid for 3-4 sec, picked out again, and then dried well. The well dried TLC was heated with a drier or on a hot plate until color was developed.
  • An external cream formulation was formulated with the composition of the following Table 8.
  • a moisturizer was added to purified water, and the resulting solution was heated and adjusted to 70° C. to form aqueous phase.
  • a fermented olive oil and other oil ingredients were heated and dissolved, and an emulsifier, a preservative and the like were then added thereto and the temperature was adjusted to 70° C.
  • the resulting solution was added to the above aqueous phase and emulsified particles were homogenized with a homomixer, and then deaeration, filtration, and cooling were performed.
  • An external lotion formulation was formulated with the composition of the following Table 9.
  • a moisturizer was added to purified water, and the resulting solution was heated and adjusted to 70° C. to form aqueous phase.
  • a fermented olive oil and other oil ingredients were heated and dissolved, and an emulsifier, a preservative and the like were then added thereto and the temperature was adjusted to 70° C.
  • the resulting solution was added to the above aqueous phase and homogenized with a homomixer, hyaluronic acid aqueous solution was added thereto and homogeneously mixed with a homomixer, and then deaeration, filtration, and cooling were performed.

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CN120168369A (zh) * 2025-05-14 2025-06-20 中山中研化妆品有限公司 一种半连续补料发酵工艺生产发酵植物油的方法

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